LOCUS       Exported                7753 bp ds-DNA     circular SYN 13-MAY-2021
DEFINITION  Yeast Expression plasmid for SARS-CoV-2 nsp2.
ACCESSION   .
VERSION     .
KEYWORDS    pGBW-m4046375
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7753)
  TITLE     SARS-CoV-2 protein Yeast expression constructs, tagged, for protein 
            expression and/or purification
REFERENCE   2  (bases 1 to 7753)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported May 13, 2021 from SnapGene Server 1.1.58
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..7753
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     promoter        10..417
                     /gene="S. cerevisiae TEF1"
                     /label=TEF1 promoter
                     /note="promoter for EF-1-alpha"
     CDS             2348..2365
                     /codon_start=1
                     /product="thrombin recognition and cleavage site"
                     /label=thrombin site
                     /translation="LVPRGS"
     primer_bind     complement(3508..3526)
                     /label=CYC1
                     /note="CYC1 transcription termination signal, reverse 
                     primer"
     CDS             3877..4680
                     /codon_start=1
                     /gene="S. cerevisiae URA3"
                     /product="orotidine-5'-phosphate decarboxylase, required 
                     for uracil biosynthesis"
                     /label=URA3
                     /note="yeast auxotrophic marker, counterselectable with 
                     5-fluoroorotic acid (5-FOA)"
                     /translation="MSKATYKERAATHPSPVAAKLFNIMHEKQTNLCASLDVRTTKELL
                     ELVEALGPKICLLKTHVDILTDFSMEGTVKPLKALSAKYNFLLFEDRKFADIGNTVKLQ
                     YSAGVYRIAEWADITNAHGVVGPGIVSGLKQAAEEVTKEPRGLLMLAELSCKGSLSTGE
                     YTKGTVDIAKSDKDFVIGFIAQRDMGGRDEGYDWLIMTPGVGLDDKGDALGQQYRTVDD
                     VVSTGSDIIIVGRGLFAKGRDAKVEGERYRKAGWEAYLRRCGQQN"
     rep_origin      4763..5745
                     /label=2u ori
                     /note="yeast 2u plasmid origin of replication"
     terminator      5823..5852
                     /label=T3Te terminator
                     /note="phage T3 early transcription terminator"
     rep_origin      5874..6461
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     terminator      6473..6500
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     CDS             complement(6524..6715)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(6716..7576)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESLRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLATIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGSLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     7339..7358
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7577..7679)
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     terminator      7680..7711
                     /label=tonB terminator
                     /note="bidirectional E. coli tonB-P14 transcription 
                     terminator"
ORIGIN
        1 gcggccgccc cacacaccat agcttcaaaa tgtttctact ccttttttac tcttccagat
       61 tttctcggac tccgcgcatc gccgtaccac ttcaaaacac ccaagcacag catactaaat
      121 ttcccctctt tcttcctcta gggtgtcgtt aattacccgt actaaaggtt tggaaaagaa
      181 aaaagagacc gcctcgtttc tttttcttcg tcgaaaaagg caataaaaat ttttatcacg
      241 tttctttttc ttgaaaattt ttttttttga tttttttctc tttcgatgac ctcccattga
      301 tatttaagtt aataaacggt cttcaatttc tcaagtttca gtttcatttt tcttgttcta
      361 ttacaacttt ttttacttct tgctcattag aaagaaagca tagcaatcta atctaagttt
      421 taattacaaa atggcataca cgaggtatgt agacaataac ttttgcggtc ctgatggata
      481 cccactggaa tgtatcaaag atttgttagc tcgtgccggt aaggctagct gtaccctttc
      541 tgagcaattg gacttcatag acactaagag aggtgtttac tgttgtcgcg aacatgaaca
      601 cgaaattgcc tggtataccg agagatcaga aaaatcctac gaattacaga ctccattcga
      661 aattaagttg gctaagaagt ttgatacatt caacggcgaa tgccccaatt tcgtctttcc
      721 attgaactct atcattaaaa ctatccaacc gagagtcgag aagaagaaat tggatggttt
      781 tatgggtaga attagatcgg tgtacccagt tgcttcccca aatgaatgta accaaatgtg
      841 tctatctact ttgatgaagt gtgaccactg tggtgaaacc agttggcaaa caggcgactt
      901 cgttaaggct acatgtgaat tttgcggtac cgagaacctc actaaggaag gtgcgactac
      961 ttgtggttat ttacctcaaa atgcagttgt caagatttac tgtccagctt gccataactc
     1021 tgaagttgga cctgaacaca gcttggctga ataccacaac gaatccggtt taaagaccat
     1081 ccttcgaaaa ggtggtcgta ccatagcctt cgggggttgc gtcttctcat atgttggttg
     1141 tcataataag tgtgcctatt gggttccaag agcttctgct aatatcggat gtaaccacac
     1201 gggtgtggtc ggtgaaggct ctgagggttt gaacgataac ttgttggaaa tacttcagaa
     1261 ggaaaaggtc aacattaaca tcgttggtga ctttaaattg aatgaagaaa ttgctattat
     1321 cctagcgtcc ttctcggcct caacttccgc ttttgtagaa actgttaagg gtcttgatta
     1381 caaagctttc aagcaaatcg tagaatcatg cggtaatttc aaggttacca aggggaaagc
     1441 caaaaaaggt gcctggaaca ttggtgaaca aaagtctata ctgagtccat tatatgcttt
     1501 cgcatctgaa gctgctagag tcgtcagatc tattttttcc agaactctcg aaaccgctca
     1561 aaattcagtg agagttttgc aaaaagcagc cattactatc ttggacggta tttcccagta
     1621 ctctttgagg ttaatcgatg ctatgatgtt tacctccgac ttggctacta ataacctggt
     1681 cgtcatggct tacataaccg gtggcgttgt tcaattgaca tctcaatggc taactaatat
     1741 attcggtact gtttatgaaa agctcaagcc agtcttggat tggttggagg aaaagttcaa
     1801 ggagggcgtt gagtttttac gggatggttg ggaaatcgtt aagttcattt ctacatgtgc
     1861 ctgtgaaatt gtcggtggtc aaatcgtgac ctgtgctaag gaaattaagg agtcggttca
     1921 aactttcttt aagttggtga acaaattttt agctctctgt gctgattcga ttatcatcgg
     1981 aggagcgaaa ttgaaggcac tgaacttggg tgaaaccttc gtcacccact ccaaaggtct
     2041 gtaccgtaaa tgtgttaaat caagagaaga aactggtttg ctaatgcctt tgaaggcccc
     2101 gaaggaaatt atcttcttgg aaggtgaaac attaccaaca gaggttttaa cggaagaggt
     2161 agttttgaag acgggggatt tgcagccgtt agaacaacca acctctgaag ccgtggaagc
     2221 tcccctagtt ggaacgccag tttgcattaa cggtctgatg ttattggaaa tcaaggacac
     2281 cgaaaagtac tgtgctctcg ctccaaacat gatggtcact aacaatacct tcactttgaa
     2341 gggtggtctc gtgccgagag gttctaaaat cgaggaaggt aagctagtca tttggataaa
     2401 tggagataag ggttacaacg gtttggcaga agtaggcaag aaatttgaaa aggacacagg
     2461 tatcaaggtg acggttgaac atccagacaa gttagaagaa aaattccctc aagttgctgc
     2521 tactggtgat ggtcccgaca ttattttctg ggcccacgat cgttttggtg gatatgctca
     2581 gtcaggtttg ctggctgaga tcaccccaga caaggccttc caagataaat tgtacccatt
     2641 cacttgggat gctgtcagat acaacggtaa gcttatagca tatccaattg ctgttgaagc
     2701 cttatcgttg atctacaata aggacttgtt acctaaccca ccaaagacat gggaagaaat
     2761 tccagctttg gataaagagc taaaggccaa gggaaaaagc gctttgatgt ttaacttaca
     2821 agaaccttat ttcacctggc cattaatcgc tgctgacggt ggttacgcat ttaaatacga
     2881 aaacggtaag tatgacatta aggatgtcgg cgttgataat gctggtgcaa aggctggttt
     2941 gaccttcttg gttgatttaa taaagaataa acacatgaac gctgacactg actactctat
     3001 cgccgaagcc gctttcaaca agggtgaaac tgctatgacc ataaatggtc catgggcctg
     3061 gagtaacatt gatacttcta aagtaaacta tggtgtcact gttttgccta cattcaaagg
     3121 ccaaccatcc aagccctttg tcggtgttct atccgctggc atcaatgctg cttctccaaa
     3181 caaggagcta gctaaagaat ttttggaaaa ttacttgtta actgacgaag gtttggaggc
     3241 cgttaacaag gacaagccat tgggtgctgt cgctttgaaa tcttacgaag aagaattggc
     3301 taaggatccc agaattgccg ctaccatgga gaacgcccaa aagggtgaaa ttatgccaaa
     3361 catcccacaa atgtcagcct tctggtacgc agttaggact gccgtgatta atgctgcttc
     3421 cggtagacaa accgtcgacg aagcattaaa agatgctcaa acgtaaacag gccccttttc
     3481 ctttgtcgat atcatgtaat tagttatgtc acgcttacat tcacgccctc ctcccacatc
     3541 cgctctaacc gaaaaggaag gagttagaca acctgaagtc taggtcccta tttatttttt
     3601 ttaatagtta tgttagtatt aagaacgtta tttatatttc aaatttttct tttttttctg
     3661 tacaaagcgg ccgccgaacc actagtgcac tgcagtacac ggtttccttg aaattttttt
     3721 gattcggtaa tctccgaaca gaaggaagaa cgaaggaagg agcacagact tagattggta
     3781 tatatacgca tatgtagtgt tgaagaaaca tgaaattgcc cagtattctt aacccaactg
     3841 cacagaacaa aaacgtgcag gaaacgaaga taaatcatgt cgaaagctac atataaggaa
     3901 cgtgctgcta ctcatcctag tcctgttgct gccaagctat ttaatatcat gcacgaaaag
     3961 caaacaaact tgtgtgcttc attggatgtt cgtaccacca aggaattact ggagttagtt
     4021 gaagcattag gtcccaaaat ttgtttacta aaaacacatg tggatatctt gactgatttt
     4081 tccatggagg gcacagttaa gccgctaaag gcattatccg ccaagtacaa ttttttactc
     4141 ttcgaggaca gaaaatttgc tgacattggt aatacagtca aattgcagta ctctgcgggt
     4201 gtatacagaa tagcagaatg ggcagacatt acgaatgcac acggtgtggt gggcccaggt
     4261 attgttagcg gtttgaagca ggcggcagaa gaagtaacaa aggaacctag aggccttttg
     4321 atgttagcag aattgtcatg caagggctcc ctatctactg gagaatatac taagggtact
     4381 gttgacattg cgaagagcga caaagatttt gttatcggct ttattgctca aagagacatg
     4441 ggtggaagag atgaaggtta cgattggttg attatgacac ccggtgtggg tttagatgac
     4501 aagggagatg cattgggtca acagtataga accgtggatg atgtggtttc tacaggatct
     4561 gacattatta ttgttggaag aggactattt gcaaagggaa gggatgctaa ggtagagggt
     4621 gaacgttaca gaaaagcagg ctgggaagca tatttgagaa gatgcggcca gcaaaactaa
     4681 aaaactgtat tataagtaaa tgcatgtata ctaaactcac aaattagagc ttcaatttaa
     4741 ttatatcagt tattacccga gtaacgaagc atctgtgctt cattttgtag aacaaaaatg
     4801 caacgcgaga gcgctaattt ttcaaacaaa gaatctgagc tgcattttta cagaacagaa
     4861 atgcaacgcg aaagcgctat tttaccaacg aagaatctgt gcttcatttt tgtaaaacaa
     4921 aaatgcaacg cgagagcgct aatttttcaa acaaagaatc tgagctgcat ttttacagaa
     4981 cagaaatgca acgcgagagc gctattttac caacaaagaa tctatacttc ttttttgttc
     5041 tacaaaaatg catcccgaga gcgctatttt tctaacaaag catcttagat tacttttttt
     5101 ctcctttgtg cgctctataa tgcagtctct tgataacttt ttgcactgta ggtccgttaa
     5161 ggttagaaga aggctacttt ggtgtctatt ttctcttcca taaaaaaagc ctgactccac
     5221 ttcccgcgtt tactgattac tagcgaagct gcgggtgcat tttttcaaga taaaggcatc
     5281 cccgattata ttctataccg atgtggattg cgcatacttt gtgaacagaa agtgatagcg
     5341 ttgatgattc ttcattggtc agaaaattat gaacggtttc ttctattttg tctctatata
     5401 ctacgtatag gaaatgttta cattttcgta ttgttttcga ttcactctat gaatagttct
     5461 tactacaatt tttttgtcta aagagtaata ctagagataa acataaaaaa tgtagaggtc
     5521 gagtttagat gcaagttcaa ggagcgaaag gtggatgggt aggttatata gggatatagc
     5581 acagagatat atagcaaaga gatacttttg agcaatgttt gtggaagcgg tattcgcaat
     5641 attttagtag ctcgttacag tccggtgcgt ttttggtttt ttgaaagtgc gtcatcagag
     5701 cgcttttggt tttcaaaagc gctctgaagt tcctatactt tctagctaga gaataggaac
     5761 ttcccgagcg gccgcgtgtt acaagcttga ctccagcgta actggactgc aatcaactca
     5821 ctggctcacc ttcacgggtg ggcctttctt cggtagaaaa tcaaaggatc ttcttgagat
     5881 cctttttttc tgcgcgtaat ctgctgcttg caaacaaaaa aaccaccgct accagcggtg
     5941 gtttgtttgc cggatcaaga gctaccaact ctttttccga ggtaactggc ttcagcagag
     6001 cgcagatacc aaatactgtt cttctagtgt agccgtagtt aggccaccac ttcaagaact
     6061 ctgtagcacc gcctacatac ctcgctctgc taatcctgtt accagtggct gctgccagtg
     6121 gcgataagtc gtgtcttacc gggttggact caagacgata gttaccggat aaggcgcagc
     6181 ggtcgggctg aacggggggt tcgtgcacac agcccagctt ggagcgaacg acctacaccg
     6241 aactgagata cctacagcgt gagctatgag aaagcgccac gcttcccgaa gggagaaagg
     6301 cggacaggta tccggtaagc ggcagggtcg gaacaggaga gcgcacgagg gagcttccag
     6361 gggtaaacgc ctggtatctt tatagtcctg tcgggtttcg ccacctctga cttgagcatc
     6421 gatttttgtg atgctcgtca ggggggcgga gcctatggaa aaacgccagc aacgcagaaa
     6481 ggcccacccg aaggtgagcc aggtgattac atttgggccc tcatcagagg ttttcaccgt
     6541 catcaccgaa acgcgcgagg cagctgcggt aaagctcatc agcgtggtcg tgaagcgatt
     6601 cacagatgtc tgcctgttca tccgcgtcca gctcgttgag tttctccaga agcgttaatg
     6661 tctggcttct gataaagcgg gccatgttaa gggcggtttt ttcctgtttg gtcacttacc
     6721 aatgcttaat cagtgaggca cctatctcag cgatctgtct atttcgttca tccatagttg
     6781 cctgactccc cgtcgtgtag ataactacga tgcgggaggg cttaccatct ggccccagtg
     6841 ctgcaatgat accgcgagac ccacgctcac cggctccaga tttatcagca ataaaccagc
     6901 cagccgggag ggccgagcgc agaagtgatc ctgcaacttt atccgcctcc atccagtcta
     6961 ttaattgttg ccgggaagct agagtaagta gttcgccagt taatagtttg cgcaacgttg
     7021 ttgccattgc tacaggcatc gtggtgtcac gctcgtcgtt tggtatggct tcattcagct
     7081 ccggttccca acgatcaagg cgagttacat gatcccccat gttgtgcaaa aaagcggtta
     7141 gctccttcgg tcctccgatc gttgccagaa gtaagttggc cgcagtgtta tcactcatgg
     7201 ttatggcagc actgcataat tctcttactg tcatgccatc cgtgagatgc ttttctgtga
     7261 ctggtgagta ctcaaccaag tcattctgag aatagtgtat gcggcgaccg agttgctctt
     7321 gcccggcgtc aatacgggat aataccgcgc cacatagcag aactttaaaa gtgctcatca
     7381 ttggaaaacg ttcttcgggg cgtaaactct caaggatctt accgctgttg agatccagtt
     7441 cgatgtaacc cactcgtgca cccaactgat cttcagcatc ttttactttc accagcgttt
     7501 ctgggtgagc aaaaacagga aggcaaaatg ccgcaaaaaa gggaataagg gcgacacgga
     7561 aatgttgaat actcatttta gcttccttag ctcctgaaaa tctcgataac tcaaaaaata
     7621 cgcccggtag tgatcttatt tcattatggt gaaagttgga acctcttacg tgccgatcaa
     7681 gtcaaaagcc tccggtcgga ggcttttgac tttctgctat ggaggtcagg tatgatttaa
     7741 atggtcagtt aga
//
